Objectives : This research was performed to investigate protective effect of Sophora Subprostrata against transient global ischemic damage after 5-min two vessel occlusion. Methods : Gerbils were divided into three groups: Normal group, 5-min two vessel occlusion (2VO) group, Sophora Subprostrata administrated group after 2VO. The CCAs were occluded by microclip for 5min. Sophora Subprostrata was administrated orally(12mg/ml) for 7 days after 2VO. The histological and immunohistochemistrical analysis was performed at 72 hours and 7 days after the surgery each. For histological analysis, the brain tissue was stained with 1% cresyl violet solution and Immunohistochemistry for BAX and Bcl-2 was carried out to examine effect of Sophora Subprostrata on ischemic brain tissue. Results : The results showed that (1) Sophora Subprostrata has the protective effect against ischemia in CA1 area of the gerbil hippocampus 7 days after 5-minute occlusion, (2) the treatment of Sophora Subprostrata inhibits the expression of Bax relatively after 2VO-induced ischemia. That protective effect of the Sophora Subprostrata seems to be performed by regulating the proportion of Bax and Bcl-2 protein, (3) in hypoxia/reperfusion model using PC12 cell, the Sophora Subprostrata extract has the protective effect against ischemia in the dose of and .This study suggests that Sophora Subprostrata has neuroprotective effect against neuronal damage following cerebral ischemia in vivo with a widely used experimental model of cerebral ischemia in Mongolian gerbils and that Sophora Subprostrata regulates the proportion of Bax and Bcl-2 protein following ischemia. And, Sophora Subprostrata extract has protective effects also on a hypoxia/reperfusion cell culture model using PC12 cell. Conclusions : Sophora Subprostrata has protective effects against ischemic brain damage at the early stage of ischemia.

SunBangHwalMyungEum (SBH) has the effects of subduing swelling, resolving masses and alleviating pain in traditional oriental medicine. Recent studies showed that SunBangHwalMyungEum produced anti-cancer, anti-metastasis and immuno-modulatory effects. However there is lack of studies regarding the effects of SBH on the immunological activities. The present study was conducted to evaluate the effect of SBH on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Methods : After the treatment of SBH, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results : Results provided evidence that SBH inhibited the production of NO, iNOS, (), IL-6, and the activation of phospholylation of inhibitor in Raw 264.7 cells activated with lipopolysaccharide. Conclusions : These findings suggest that SBH can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Thujae Semen(TSW) in PC12 cells. Methods : We performed 2,2-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging assay, 2,2-azinobis- (3-ethyl-benzothiazoline-6-sulfonic acid(ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of TSW. We also carried out 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay(MTT), reactve oxygen species(ROS) assay, and nitric oxide(NO) assay to examine neuroprotective effects against 6-hydroxydopamine(6-OHDA) in PC12 cells. Results : TSW showed values of 404.3 and 219.9 in DPPH and in ABTS assays, respectively. TSW showed 9.74 of total polyphenol contents. TSW incresed cell viability in a dose dependent manner and it showed protective effect against 6-OHDA neurotoxicity at the concentration of 25-200 . Moreover, it recovered 6-OHDA induced cell death at the same concentrations. The extract showed a dose dependent reduction of ROS and NO generation by 6-OHDA. Conclusions : We concluded that TSW has neuroprotective effect against 6-OHDA-induced toxicity in PC12 cells through ROS and NO inhibition.

Objectives : In this paper, we describe that spectroscopy may be superior to the conventional HPLC for the quantitative analysis of hesperidin from Citrus unshiu. Methods : spectra (400 MHz) were recorded in using a Varian UNITY Inova AS 400 FT NMR spectrometer. One hundred milligram of powdered Citrus unshiu was weighed out and mixed with 1 ml of with sonication for 30 min (room temperature). The extracts were filtrated through a 0.45 PVDF filter and 0.5 ml of filtrated extract used for quantitative measurement (added 1 mg of dimethyl terephthalate as internal standard). The quantity of hesperidin was calculated by the ratio of the intensity of the compound to the known amount of internal standard. For HPLC analysis, the half gram of plant material was extracted with 60 ml of MeOH for 2 hours. The extracts were made 100 ml volume and analyzed by a Waters HPLC system using a YMC ODS column. The total flow rate was 1.0 ml/min with a sample volume 10 and UV detection at 280nm. Results : The contents of hesperidin in Citrus unshiu was determined % in the quantitative method and in HPLC method. Using the quantitative the contents of hesperidin can be determined in much shorter time than the conventional HPLC measurements. Conclusions : From those results, the advantages of quantitative analysis are that can be analyzed to identify and quantify, and no reference compounds required for calibration curve. Besides, it allows rapid and simple quantification for hesperidin with an analysis time for only 10 min without any pre-purification steps.

Objectives : For the purpose of developing Korean herbalogy of the plants belonging to Pteridaceae in Korea, the literatures of the successive generations have been thoroughly investigated to prepare this article. Methods : The examined herbalogical books and research papers which published at home and abroad. Results : 1: There are totaled to 12 genera and 33 species in Pteridaceae in Korea and among them medicinal plants are 10 genera, 19 species, some 58% in total. 2: Adiantum genus is main kind enough that it has 5 species among 33 species in Pteridaceae, of which medicinal plants are 4 species. 3: The herb is the main medicinal parts if medicinal plants in the Pteridaceae, which is used in 18 species. 4: According to nature and flavor of medicinal plants in the Pteridaceae, they were classified into cold 15 species, and cool 11; bitter taste 12, little bitter taste 11 and sweet taste 5 in the order. 5: According to meridian propism of medicinal plants in the Pteridaceae, they were classified into liver meridian 15 species, lung and large intestine meridian 8 species, heart meridian 6 species. 6: According to the properties and principal curative action, they were classified into drugs for clearing away heat 24 species, drugs for detoxification 22 species, drugs for promoting diuresis 12 species in the order. 7: The number of toxic species in the Pteridaceae was examined to be 2 species. Conclusions : There are totaled to 12 genera and 33 species in Pteridaceae in Korea and among them medicinal plants are 10 genera, 19 species, some 58% in total.

Objectives : This study was conducted to evaluate the antioxidative and anticancer activity of extracts from Scutellaria baicalensis, Zizyphus jujuba and Atractylodes macrocephala. Methods : Three kinds of medicinal herbs were extracted with distilled water and 70% ethanol, and the extracts were tested for their antioxidant activity and growth inhibition activity against cancer cells. Results : Electron donating abilities (EDA) of the water and ethanol extract from Zizyphus jujuba and Atractylodes macrocephala were over than 87% at 1,000 , respectively. The highest nitrite scavenging abilities (NSA) of the water and ethanol extracts from Scutellaria baicalensis were 68.9 and 79.2% at 1,000 , respectively. Atractylodes macrocephala extracts had shown the highest growth inhibition activity against both of MDA and A549 cells and the water extract had higher activity than ethanol extract. Conclusions : These results suggest that the extracts of medicinal herbs used for this experiment had specific biological activity and they can be used as natural antioxidant to prevent oxidative damage in normal cells.

Objecives : Aucklandiae Radix is a root of Aucklandia lappa which has been widely used for regulating the flow of vital energy, invigorating the spleen, alleviating pain. Aucklandiae Radix contains the costunolide which is the main ingredient. The substitutive Aucklandiae Radix are Inulae helenii Radix, Aristolchiae Radix, Vladimiriae Radix, and Inulae racemosi Radix in Korea and China. This paper is analysised and compared the costunolide and HPLC pattern in Aucklandiae Radix and substitute herbs. Methods : Chromatographic separation performed using C18 column(Luna 5 u, 250 mm 4.6 mm) with a mixture of methanol and water(65:35)(v/v). The analyses detected at UV(210 nm). Results : Optimal extraction condition of costunolide was 100% methanol for 2hr. Costunolide was detected in Aucklandiae Radix and Vladimiriae Radix, but other herbs were not detected. In Korea herbal market, Aristolchiae Radix merchandise was identified as the imported Inulae helenii Radix. Conclusions : According to above results, this method was useful identified to Aucklandiae Radix and substitutive herbs. In Korea herbal market, Aristolchiae Radix was identified as Inulae helenii Radix.

Objectives : This study presents a high performance liquid chromatography methods for the quantitative and qualitative analysis of rosmarinic acid (RA) and caffeic acid (CA) in Perilla frutescens var. japonica and var. acuta. Methods : Chromatographic separation was performed using a mixture of methanol, water and formic acid (35 : 64.2 : 0.8) with a reversed-phase column (Gemini C18, 4.6 150 mm, 3 ). The analyses were detected at UV (280 nm). Results : The samples were extracted with 50% EtOH under reflux for 1 h, and simultaneous determination for RA and CA in hyang-so-san and haeng-so-san was possible without interference peaks Conclusions : According the results, we developed a determination method for RA and CA in Perillae Folium. Owing to Perilla frutescens var. japonica and var. acuta did not show significant difference in contents of RA and CA, both Perilla frutescens could be available as herbal medicine.

Objectives : The root of Zingiber officinale ROSC. (Zingiberis Rhizoma Recens; Ginger) has been widely used as one of folk remedies and food materials in many traditional preparations. Ginger is known as an effective appetite enhancer and anti-inflammatory agent. This study was performed to investigate the effect of ginger chloroform fraction (GCF) in microglia which play a central role on brain inflammation in neurodegenerative diseases. Methods : Dried ginger was extracted with 80% methanol, and then fractionated with chloroform. BV2 mouse microglial cells were cultured with different concentrations of GCF and then stimulated with LPS (1 ) at indicated times. The cell toxicity of GCF was determined by MTT assay. The concentrations of NO, PGE2 and cytokines were measured by Griess assay and enzyme-linked immunosorbant assay. The mRNA and protein expressions of iNOS, COX-2 and cytokines were determined by RT-PCR and Western blotting. The phosphorylation of three MAPKs (p38 MAPK, ERK1/2 and JNK) and activation were determined by Western blotting. Results : GCF significantly inhibited LPS-induced production of inflammatory mediators, NO, and proinflammatory cytokines ( and ) in a dose-dependent manner. GCF attenuated LPS-induced expression of mRNA and protein of inflammatory enzymes, iNOS, COX-2 and proinflammatory cytokines through suppressing the phosphorylation of ERK1/2 and p38 MAPK and the activation of p65 in BV2 cells. Conclusions : This study suggests that GCF may have an anti-inflammatory property through suppressing the inflammatory mediator production released by activated microglia after the brain injury.

Objectives : Forsythiae Fructus (Forsythia koreana Nakai) has been used anti-inflammatory, diuretics, antidote, and antibacterials in traditional herbal medicine. The present study is focused on the inhibitory effect of Forsythiae Fructus ethanol extract (FF-E) on the production of inflammatory mediators such as NO, iNOS and proinflammatory cytokines (, and IL-6) in LPS-stimulated BV-2 cells, a mouse microglial cell line, and investigated the scavenging activity of FF-E. Methods : BV-2 cells were pre-incubated with FF-E for 30 min and then stimulated with LPS (1 ) at indicated times. Cell toxicity of GCF was determined by MTT assay. The levels of NO, PGE2 and cytokines were measured by Griess assay and ELISA. The mRNA and protein expressions of iNOS and cytokines were determined by RT-PCR and Western blotting. Free radical scavenging activity of GCF was determined by DPPH assay in tube test. Results : FF-E significantly inhibited the excessive production of NO, , , and in LPS-stimulated BV-2 cells. In addition, FF-E attenuated the mRNA and protein expressions of iNOS, and proinflammatory cytokines. FF-E also significantly scavenged the DPPH free radicals in a dose-dependent manner. Conclusions : These results indicate that FF-E exhibits anti-inflammatory property by suppressing the transcription of inflammatory mediator genes, suggesting the anti-inflammatory property of FF-E may make it useful as a therapeutic agent for the treatment of human neurodegenerative diseases.

Objectives : This research aimed to study the cytotoxicity and immuno modulating activity of fermented Artemisia argyi Lev. et Vant.(Compositae). Methods : Effect of fermented Artemisiae Argyi Folium extracts, which were fermented by Sacchromyces cerevisiae STV89(AFS), on cell viability, generation of ROS within cells, generation of NO and the level of cytokines( and IL-6) was measured using mouse macrophage RAW 264.7 cell. Results : 1. Result of MTT assay conducted to verify the cytotoxicity of fermented Artemisiae argyi folium extract illustrated that, when fermented Artemisiae argyi folium extract was processed for each concentration, there was no excessive induction of cytoxicity in the RAW 264.7 cell. 2. Fermented Artemisiae Argyi Folium extract increased the generation of H2O2 within RAW 264.7 cell as well as significantly increased inhibition of generation of H2O2 in macrophage induced by LPS. 3. Fermented Artemisiae Argyi Folium extract inhibited generation of NO in RAW 264.7 cell, and significantly inhibited increase in generation of NO of macrophage induced by LPS. 4. Fermented Artemisiae Argyi Folium extract, AFS has significantly reduced the increase in the generation of above 10 . 5. Fermented Artemisiae Argyi Folium extract, AFS has significantly reduced the increase in generation of IL-6 above 50 . Conclusions : AFS fermented extract produced from Artemisiae Argyi Flium, have increased generation of ROS and reduced generation of NO in RAW 264.7 cell without excessively inducing cytotoxicity of RAW 264.7 cell. In addition, they displayed significant immuno modulating activities including inhibition of generation of and IL-6 in macrophage, induced by LPS.

Objectives : The purpose of this paper was to investigate the anti-inflammatory effects of extract from Teucrium veronicoides (TV) on the RAW 264.7 cells. Methods : To evaluate of anti-inflammatory of TV, we examined the cytokine productions on lipopolysacchride (LPS)-induced RAW 264.7 cells and also inhibitory mechanisms using Western blot. Furthermore, We examined LPS-induced endotoxin shock. Results : 1. Extract from TV itself does not have any cytotoxic effect. 2. Extract from TV reduced LPS-induced Nitric oxide (NO),interleukin (IL)-1b, IL-6 and IL-10, tumor necrosis factor-a (TNF-a) production in RAW 264.7 cells. 3. TV inhibited the activation of mitogen-activated protein kinases (MAPKs) such as p38, extracelluar signal-regulated kinase (ERK 1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of inhibitory kappa B a (Ik-Ba) in the LPS-stimulated RAW 264.7 cells. 3. TV slightly increased the duration of survival after LPS-induced endotoxin shock. Conclusions : TV down-regulated LPS-induced NO and cytokines production, which could provide a clinical basis for anti-inflammatory properties of TV.

Objectives : The present study was designed to investigate whether Ostericum koreanum (OK) could regulate lipopolysaccharide (LPS)-induced inflammatory response in vitro and in vivo. Methods : To evaluate of anti-inflammatory effect of OK, we examined Nitric oxide (NO), proinflammatory cytokines production in LPS-stimulated RAW264.7 cells. Furthermore, we checked molecular mechanism especially in the phosphorylation of mitogen-activated protein kinases (MAPKs) and the degradation of inhibitory kappa B a () using western blot and also investigated survival of mice in LPS-mediated endotoxin shock. Results : 1. Extract from OK itself have weak cytotoxic effect on RAW264.7 cells. Extract from OK inhibited LPS-induced NO, tumor necrosis (), interleukin , IL-6 and IL-10 production in RAW264.7 cells. 2. OK inhibited the phosphorylation of MAPKs, such as p38, extracelluar signal-regulated kinase (ERK1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of in the LPS-stimulated RAW264.7 cells 3. OK did not inhibit LPS-induced endotoxin shock. Conclusions : OK down-regulated LPS-induced NO and cytokines production through suppressing activation of MAPKs and degradation of . Our results suggested that OK may be a beneficial drug against inflammatory diseases.

Objectives : The purpose of this study is to clarify the medicinal using part of Linderae Radix. Methods : Find out the using part of Linderae Radix in the Pharmacopoeia, main Herbal book and present day scientific research result. Results : 1. In the Phamacopoeia of 6 Nation, The herbal name of Lindera strychnifolia is Linderae Radix, the origin is root, shapes of Linderae Radix are recorded spindle-shaped, slightly curved, some constricted in the middle to be moniliform on every Pharmacopoeia. But only in the Pharmacopoeoa of the people's republic of china, root is recorded as root tuber. 2. The Straight root of Linderae Radix can not be used as medicine on the Pharmacopoeia of the China, Vietnamese, Chinese Taipei. 3. In Gaebo Boncho(開寶本草) and Bonchodogyeong(本草圖經), Dongeuibogam(東醫寶鑑), Bonchogangmok(本草綱目), ickmulmyeongsildogo(植物名實圖考), only the spindle-shaped root of Linderae Radix can be used as medicine. 5. The picture of the root is also spindle-shaped. Conclusions : The herbal name of Lindera strychnifolia is Linderae Radix, medicinal using part is root tuber, spindle-shaped, some constricted in the middle to be moniliform, straight and not spindle-shaped tap roots can not be used medicinally, and allowing guide line of straight root will upgrade the quality of Linderae Radix.

Objectives : The cortex and root of Acanthopanax sessiliflorus (AR), a herbal medicine, have been used for several diseases including cancer in Oriental countries. Recently, we reported that AR has an immune-potentiating action. Oga-Power(OP) was made using extract from AR. For these reasons, we hypothesized that OP can potentiate the immune system in terms of accelerating proliferation rates of immune cells such as thymocytes and splenocytes. Methods : In this experiment, proliferation rates of thymocytes and splenocytes were measured using modified 3-[4,5-dimethy -lthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). No production levels in macrophages isolated from normal mice were measured using Griess method. Results : In our results, treatment with OP accelerated proliferation rates of splenocytes, but did not affect those of thymocytes in vitro. On the other hand, proliferation rates of thymocytes was elevated in vivo. In addition, level of NO production from macrophage separated from abdominal cavity of normal mice was elevated by treatment with OP. Conclusions : In conclusion, OP has immune-potentiating action, by acceleration of splenocyte proliferation and elevation of NO production level from macrophages.

Objectives : The purpose of this study is to investigate the biological Effect of multi-herbal drug 'KOCO-P1' on human hepatocyte HepG2 cells. Methods : Multi-herbal drug 'KOCO-P1' was composed of Ginseng Radix, Astragali Radix, Polygonati Rhizoma, Liriopis Tuber, and Scrophulariae Radix. Cytotoxicity and cytoprotective activity of KOCO-P1 was verificated by MTT assay. And antioxidative effect of KOCO-P1 against EtOH, Nicotine was inspected by Hydroperoxide assay. Results : KOCO-P1 showed no cytotoxicity on HepG2 cells for 24, 48 hours. KOCO-P1 at 50 reduced the production of H2O2 in HepG2 cells by EtOH. KOCO-P1 at 50 reduced the production of in cells by Nicotine. Conclusions : KOCO-P1 at the low concentration could be supposed to have antioxidative effect on human hepatocyte with no cytotoxicity.

Objectives : This study was evaluated to elucidate the inhibitory potential of Bodusan (BDS) and its components, Strychnos ignatii semen (SIS) and Glycyrrhizae Radix (GR), on human aortic smooth muscle cells (HASMC) migration and production of matrix metalloproteinase (MMP)-2 and MMP-9 by treatment. Methods : Cytotoxic activity of BDS and its components on HASMC was using 5-(3-caroboxymeth- oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Effect of BDS, SIS and GR on HASMC migration underside of matrigel filter was stained with hematoxylin-eosin. And total number of cells that migrated to the underside of the filter was counted. MMP-2 and MMP-9 activity was evaluated by gelatin zymography assay. Results : The matrigel migration assay showed that BDS effectively inhibited the migration of HASMC. Moreover, BDS significantly inhibited MMP-9 activity. Our present study demonstrates that BDS and its components inhibits HASMC migration and MMP-9 activity. The inhibitory effect of BDS extract is more potent than that of its component herb extracts. Conclusions : These results provide evidence that BDS has multiple effects in the inhibition of HASMC migration and may offer a therapeutic approach to block HASMC migration.

Objectives : Imyosan (IMS), a drug preparation comprised of Phellodendri Cortex (PC) and Atractylodis Rhizoma (AR), is commonly used as a traditional herbal medicine in Korea and China for the treatment of various inflammatory diseases. However, little is known about the effect of IMS and its component herbs on inflammatory mediators in RAW 264.7 cells. Therefore, in this study, methanol extracts of IMS and its component herbs were examined to determine if they inhibited inflammatory effects in RAW 264.7 cells. Methods : Cytotoxic activity of IJHT and its components on RAW 264.7 cells was using 5-(3-carboxymethoxyphenyl)-2H-tetrazolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : Methanol extract of IMS and its component herbs were significantly reduced iNOS and COX-2 expression as well as NO, PGE2, and IL-6 production in RAW 264.7 cells. Conclusions : The results of this study indicated that the anti-inflammatory effect of Imyosan extract is more potent than that of extracts of its component herbs in macrophages.

Objecitivies : While Glechoma longituba (Nakai) Kupr(GL) has been known as a antiinflammatory, antidiabetic and antihypertension medicine in korea traditional medicine, there has been only a few researches about it in Korea. Thus, we examined antidiabetic effect of GL in diabetic rats induced by streptozotocin Methods : To verify the effects of GL on serum glucose, creatinine, BUN, total cholesterol and triglyceride through streptozotocin-induced diabetic rats, some rats were injected 1 time with 55mg/kg streptozotocin intraperitoneally to induce diabetes, and other rats were injected with buffer I.P. as control group. Results : GL extracts significantly reduced the amount of serum glucose, BUN, creatinine, and triglyceride level in diabetics rats induced by streptozotocin. Conclusions : According to above result, Glechoma longituba was expected to be applied to the treatment of diabetes mellitus.